LOCUS       40924_735        6780 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6780)
  AUTHORS   Sun Q, Westphal W, Wong KN, Tan I, Zhong Q
  TITLE     Rubicon controls endosome maturation as a Rab7 effector.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Nov 9. 107(45):19338-43.
  PUBMED    20974968
REFERENCE   2  (bases 1 to 6780)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6780)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2010 Nov 9. 107(45):19338-43."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(258..391)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(568..1359)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    complement(2243..2467)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(3170..3196)
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             complement(3197..3223)
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             complement(3233..3949)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(3963..3981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4026..4229)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(4230..4609)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4781..4800
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     promoter        4875..4979
                     /label=AmpR promoter
     CDS             4980..5837
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6011..6599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6753..6770
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgagcg gccgcttagc aactgcagct ttctgccgag gccttggccc
     2581 ggtcattctt gtccagtttg ataggttcag gaaattcgtt gtacagctcc acctccgttt
     2641 cctgcttaag tgcattccgt gcaatcgtct ggaacgcctg ctccacgttg atggcctcct
     2701 tggcactggt ctcaaagtag ggaatgttgt ttttgctgta gcaccaggcc tgtgcccgct
     2761 ttgtggccac ttgtctgttt tcgaggtcaa tcttgtttcc caacacaaca aatgggaagt
     2821 tttcaggatc tcggggactg gcctggatga gaaactcatc tctccagcta tctagggttt
     2881 tgaatgtgtt gggggcagtc acatcaaata ccagaacgca gcagtctgca cctctgtaga
     2941 aggccacacc gagagactgg aaccgttcca gtcctgctgt gtcccatatc tgcattgtga
     3001 ctagcctgtc atccaccatc acctccttgg tcagaaagtc agctcctatt gtggctttgt
     3061 actgattgct gaatttctta ttcacatact ggttcatgag tgatgtcttc ccgactccag
     3121 aatctcccag gatgataacc ttcagcaaca ctttcttcct agaggtacca gcgtaatccg
     3181 gaacatcgta tgggtaagcg taatccggaa catcgtatgg gtacatggat cccttgtaca
     3241 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     3301 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg
     3361 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     3421 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     3481 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     3541 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     3601 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     3661 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     3721 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     3781 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     3841 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     3901 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccata agcttgggtc
     3961 tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc
     4021 cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     4081 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     4141 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     4201 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     4261 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     4321 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     4381 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     4441 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     4501 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     4561 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
     4621 ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa
     4681 ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc
     4741 tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta
     4801 ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt
     4861 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4921 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4981 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5041 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5101 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5161 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5221 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5281 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5341 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5401 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5461 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5521 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5581 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5641 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5701 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5761 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5821 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5881 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5941 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6001 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6061 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
